Fig. 1: Pp4c interacts with Smek1, CreA, and Crf1 in Magnaporthe oryzae. | Communications Biology

Fig. 1: Pp4c interacts with Smek1, CreA, and Crf1 in Magnaporthe oryzae.

From: A multilayered regulatory network mediated by protein phosphatase 4 controls carbon catabolite repression and de-repression in Magnaporthe oryzae

Fig. 1

a Yeast two-hybrid assays between Pp4c and Smek1, CreA, and Crf1. Yeast strains carrying prey and bait vectors were cultured on SD-Leu-Trp and SD-Leu/Trp/His/Ade media at 30 °C for 4 days. Strains containing pGADT7-T and pGBKT7-53 were used as positive controls. b GST pull-down assays detecting Pp4c-Smek1 interactions. GST-tagged fusion proteins were expressed in Escherichia coli BL21 and pulled down using anti-GST beads. Eluted proteins were detected with anti-GST and anti-Flag antibodies. c Co-immunoprecipitation assays to detect Pp4c interactions with Smek1 and Crf1. Total proteins extracted from strains expressing PP4C-FLAG and GFP-CRF1 or PP4C-FLAG and GFP-SMEK1 were precipitated with anti-GFP beads. Eluted proteins were detected using anti-GFP and anti-Flag antibodies. d GST pull-down assays detecting Pp4c-CreA interactions.

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